Inhibition of calcium-dependent motility of cochlear outer hair cells by the protein kinase inhibitor, ML-9.
The calcium ionophore ionomycin has been shown to induce length increases of guinea pig outer hair cells (Dulon et al., 1990). We have demonstrated that these length increases can be inhibited by a 30 min preincubation of the cells with the protein kinase inhibitor ML-9. At either 30 or 60 s after ionomycin application, the effect of ML-9 was dose-dependent with a half maximal response at approximately 0.3 microM. No effect on cell length was detected after 30 min incubation with 0.5 and 5 microM ML-9 alone. However, with 50 and 500 microM ML-9, significant contraction in cell length was observed. 50 microM ML-9 did not interfere with the ability of ionomycin to elevate fluorescence of the calcium indicator Fluo-3, nor did it alter the ability of cells to exclude propidium iodide from their nuclei. Treatment with 500 microM ML-9 resulted in impaired cell morphology. The data support the hypothesis that protein kinase activity regulates calcium-dependent processes that affect shape changes of outer hair cells. They are consistent with the involvement of the calcium/calmodulin-dependent enzyme, myosin light chain kinase, a known target of ML-9, but do not preclude the possibility of another intracellular target for ML-9.
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Kresge Hearing Research Institute, University of Michigan, Ann Arbor 48109-0506, USA. fortelab@lsa.berkeley.edu
Bartolami
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Rhee
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D000814:Aniline Compounds; D000818:Animals; D001381:Azepines / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival; D003051:Cochlea / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000187:drug effects* / Q000502:physiology; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors*; D011419:Propidium; D014966:Xanthenes
D000814:Aniline Compounds; D001381:Azepines; D004791:Enzyme Inhibitors; D007476:Ionophores; D014966:Xanthenes; C056218:ML 9; C059715:Fluo-3; D011419:Propidium; D015759:Ionomycin; D009219:Myosin-Light-Chain Kinase; D002118:Calcium